以整合素连接激酶相关磷酸酶(ILKAP)全蛋白为抗原,采用腿部肌肉注射和耳缘静脉注射相结合的方法对新西兰大白兔进行免疫,共免疫6次,最终获得ILKAP抗血清.采用Protein A agrose柱纯化抗体,获得ILKAP的多克隆抗体.免疫印迹分析表明,所获得...以整合素连接激酶相关磷酸酶(ILKAP)全蛋白为抗原,采用腿部肌肉注射和耳缘静脉注射相结合的方法对新西兰大白兔进行免疫,共免疫6次,最终获得ILKAP抗血清.采用Protein A agrose柱纯化抗体,获得ILKAP的多克隆抗体.免疫印迹分析表明,所获得的ILKAP多克隆抗体具有较高的特异性,间接ELISA方法测定ILKAP多克隆抗体的滴度为1∶1 600.免疫荧光分析表明,所获得的ILKAP多克隆抗体在细胞水平应用效果较好.展开更多
Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality,necessitating novel therapeutic targets.This study explores the oncogenic role of integrin-linked kinase-associated phosphatase(ILKAP)in HCC ...Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality,necessitating novel therapeutic targets.This study explores the oncogenic role of integrin-linked kinase-associated phosphatase(ILKAP)in HCC and its underlying mechanisms.Database analyses(TCGA,UALCAN)revealed ILKAP overexpression in HCC,correlating with poor prognosis.Functional assays demonstrated that ILKAP knockdown significantly suppressed HCC cell proliferation and migration in vitro,while xenograft models confirmed its role in tumor growth in vivo.RNA sequencing identified 357 differentially expressed genes(DEGs),including 48 protein-coding DEGs,with glycolytic enzyme PGAM1 notably downregulated upon ILKAP silencing.ILKAP and PGAM1 expression were positively correlated in HCC tissues,and elevated PGAM1 levels were linked to worse survival.Notably,restoring PGAM1 in ILKAP-knockdown cells rescued proliferation and invasion,underscoring PGAM1’s critical role in ILKAP-mediated tumor progression.ILKAP depletion also reduced extracellular acidification rates and altered glycolysis-related gene expression,highlighting its role in metabolic reprogramming.These findings suggest that ILKAP drives HCC malignancy by modulating PGAM1 and glycolysis,providing a potential therapeutic target for HCC treatment.Further elucidation of the ILKAP-PGAM1 axis may offer new strategies for liver cancer management.展开更多
文摘以整合素连接激酶相关磷酸酶(ILKAP)全蛋白为抗原,采用腿部肌肉注射和耳缘静脉注射相结合的方法对新西兰大白兔进行免疫,共免疫6次,最终获得ILKAP抗血清.采用Protein A agrose柱纯化抗体,获得ILKAP的多克隆抗体.免疫印迹分析表明,所获得的ILKAP多克隆抗体具有较高的特异性,间接ELISA方法测定ILKAP多克隆抗体的滴度为1∶1 600.免疫荧光分析表明,所获得的ILKAP多克隆抗体在细胞水平应用效果较好.
基金supported by grants from the Natural Science Foundation of Zhejiang Province(No.LGF22H080016)the Key Research and Development Select Projects of Zhejiang Provincial Department of Science and Technology(No.2020C03008)+1 种基金the Zhejiang Medical Health Science and Technology Project(No.2022RC124)the Hangzhou Medical College Basal Research Fund(No.KYZD2024004).
文摘Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality,necessitating novel therapeutic targets.This study explores the oncogenic role of integrin-linked kinase-associated phosphatase(ILKAP)in HCC and its underlying mechanisms.Database analyses(TCGA,UALCAN)revealed ILKAP overexpression in HCC,correlating with poor prognosis.Functional assays demonstrated that ILKAP knockdown significantly suppressed HCC cell proliferation and migration in vitro,while xenograft models confirmed its role in tumor growth in vivo.RNA sequencing identified 357 differentially expressed genes(DEGs),including 48 protein-coding DEGs,with glycolytic enzyme PGAM1 notably downregulated upon ILKAP silencing.ILKAP and PGAM1 expression were positively correlated in HCC tissues,and elevated PGAM1 levels were linked to worse survival.Notably,restoring PGAM1 in ILKAP-knockdown cells rescued proliferation and invasion,underscoring PGAM1’s critical role in ILKAP-mediated tumor progression.ILKAP depletion also reduced extracellular acidification rates and altered glycolysis-related gene expression,highlighting its role in metabolic reprogramming.These findings suggest that ILKAP drives HCC malignancy by modulating PGAM1 and glycolysis,providing a potential therapeutic target for HCC treatment.Further elucidation of the ILKAP-PGAM1 axis may offer new strategies for liver cancer management.
